Specific binding of kainic acid to purified subcellular fractions from rat brain.

Abstract

The subcellular distribution of kainic acid (KA) binding sites in rat brain has been studied using a microcentrifugation assay. KA did not bind to myelin or brain cytosol and had few or no binding sites in the nuclear fraction. However, it bound to microsomal components (Kd = 128-136 nM; 2.5-4.8 pmol/mg protein), purified synaptic plasma membranes (SPM)(Kd = 45-71 nM; 5.8-6.5 pmol/mg), and purified cell-body and intraterminal mitochondria (Kd = 11-31 nM; 0.4-1.1 pmol/ng). Bound KA could be totally displaced by L-glutamate or L-asparate, but several putative antagonists of these amino acids (nuciferin, compound HA-966, 2-amino-4-phosphonobutyrate, and 2-amino-3-phosphonoproprionate) failed to displace KA or did so at very high concentrations (greater than or equal to 4 mM). Glutamic acid diethyl ester (GDEE) and D,L-alpha-aminoadipate (alpha-AA) were more effective (IC50, 0.2-0.8 mM) and showed differential effects in their capacity to displace KA bound to the various subcellular fractions. Thus, GDEE only displaced 40-60% of the KA bound by SPM or mitochondria and did not prevent the binding of KA to microsomes. alpha-AA, on the other hand, was more effective in preventing the binding of KA at high concentrations and displaced between 80 and 100% of the drug. Both compounds showed biphasic curves of KA displacement from synaptic plasma membranes and mitochondria. The overall results indicate the presence of multiple binding sites for KA in brain cells and suggest that KA does not act exclusively as synaptic glutamate receptors. The mechanism of KA action is most likely quite complex, and the drug probably acts at multiple binding sites affecting a number of processes.